Synthesis of Poly(Malic Acid) Derivatives End-Functionalized with Peptides and Preparation of Biocompatible Nanoparticles to Target Hepatoma Cells.

Clarisse Brossard,Catherine Ribault,Sandrine Cammas-Marion,Nicolas Noiret,Pascal Loyer,Vincent Dorcet,Elise Vène,Manuel Vlach,Nicolas Lepareur

doi:10.3390/nano11040958

Abstract

Recently, short synthetic peptides have gained interest as targeting agents in the design of site-specific nanomedicines. In this context, our work aimed at developing new tools for the diagnosis and/or therapy of hepatocellular carcinoma (HCC) by grafting the hepatotropic George Baker (GB) virus A (GBVA10-9) and Plasmodium circumsporozoite protein (CPB)-derived peptides to the biocompatible poly(benzyl malate), PMLABe. We successfully synthesized PMLABe derivatives end-functionalized with peptides GBVA10-9, CPB, and their corresponding scrambled peptides through a thiol/maleimide reaction. The corresponding nanoparticles (NPs), varying by the nature of the peptide (GBVA10-9, CPB, and their scrambled peptides) and the absence or presence of poly(ethylene glycol) were also successfully formulated using nanoprecipitation technique. NPs were further characterized by dynamic light scattering (DLS), electrophoretic light scattering (ELS) and transmission electron microscopy (TEM), highlighting a diameter lower than 150 nm, a negative surface charge, and a more or less spherical shape. Moreover, a fluorescent probe (DiD Oil) has been encapsulated during the nanoprecipitation process. Finally, preliminary in vitro internalisation assays using HepaRG hepatoma cells demonstrated that CPB peptide-functionalized PMLABe NPs were efficiently internalized by endocytosis, and that such nanoobjects may be promising drug delivery systems for the theranostics of HCC.

Highlights

At the beginning of the 20th century, Paul Ehrlich coined the term “Magic Bullet”, postulating that the targeting of drug towards specific diseased cells/tissues could be achieved without affecting healthy ones [1]
We demonstrated that the same biotinylated peptides, when bridged via the streptavidin to biotinylated nanoparticles (NPs) constituted by the amphiphilic block copolymer biotin-poly(ethylene glycol)-block-poly(benzyl malate) (BiotPEG-bcopolymer biotin-poly(ethylene glycol)-block-poly(benzyl malate) (BiotPEG-b-PMLABe), PMLABe), significantly enhanced the cell uptake by human hepatoma but nothuby significantly enhanced the cell uptake by human hepatoma cells but notcells by normal normal human hepatocytes
These results demonstrate the success of the peptide grafting onto maleimide end-functionalized PMLABe73 and PEG62 -b-PMLABe73

Summary

Introduction

At the beginning of the 20th century, Paul Ehrlich coined the term “Magic Bullet”, postulating that the targeting of drug towards specific diseased cells/tissues could be achieved without affecting healthy ones [1]. Tremendous research work has been done for designing efficient site-specific drugs using various ligand molecules, such as monoclonal antibodies (mAbs) [2,3,4], vitamins [2,5,6,7], carbohydrates [2,8], and aptamers [9,10,11]. In 1902 Theodor Curtius synthesized benzoyl-glycyl-glycine using the azide reaction he developed [17]. Following these pioneer works, researchers worked to increase the number of amino acids in the peptide’s chain, which was possible thanks to the use of the removable benzyloxycarbonyl protecting group developed by Max Bergmann and Leonidas Zervas [17]. The uses of peptides as therapeutic molecules emerged in the

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Conclusion